79 results about "Selectable marker" patented technology

The invention provides application of a pig STR molecular marker combination in pig individual identification, a kit and an identification method, relates to the technical field of biology, and develops an STR molecular marker combination for accurate identification of pig individual identification by applying a whole genome association analysis method for 18 wild boar varieties and 42 domestic pig varieties. By detecting the combination, individual identification and genetic identification can be accurately and efficiently carried out on a to-be-detected pig sample, and when the minimum set number of the selected STR marker is 6, individual distinguishing can be carried out. According to the scheme provided by the invention, by comprehensively analyzing the STR typing result, the parent-child margin determination and individual identification of the pig can be effectively carried out, so that the pedigree accuracy in pig breeding work can be improved, the breeding process is accelerated, and the method has good application prospects and economic values.

The present invention relates generally to the field of molecular biology and concerns a method for high throughput screening and characterization of one or more candidate plant pathogen effect genes and / or one or more candidate plant disease resistance genes in a single plant cell, the method comprises: (i) introducing into a plant protoplast one or more expression cassettes comprising at least one candidate plant pathogen effect gene and / or at least one candidate plant disease resistance gene; (ii) transiently expressing the one or more candidate plant pathogen effect genes and / or the one or more candidate plant disease resistance genes; (iii) measuring reactive oxygen species (ROS) produced by the protoplast; then (iv) separating the plant protoplast producing the ROS; and (v) identifying the introduced at least one candidate plant pathogen effect gene and / or at least one candidate plant disease resistance gene. Alternatively, the protoplast is capable of expressing the selectable marker, thereby allowing for the measurement of modulated selectable marker expression as an alternative indicator for HR activation.

A method of producing a population of mature human pluripotent stem cell-derived sensory neurons (hPSC-SNs) expressing a target gene associated with at least one of nociceptive pain, chronic pain, pruriception and a nociceptive- or pruriceptive- mediated condition, the method comprising introducing into a population of human pluripotent stem cells (hPSCs) a composition comprising at least one site-directed nuclease targeting a site within the target gene, and at least one nucleic acid comprising a nucleotide sequence encoding at least one screenable, selectable marker that is flanked by (i) a nucleotide sequence homologous with a region located upstream of the target site within the target gene and (ii) a nucleotide sequence homologous with a region located downstream of the target site within the target gene, wherein the target site is located downstream of the open reading frame of the target gene, and wherein the site-directed nuclease cleaves the target site of the target gene and the nucleic acid encoding the screenable, selectable marker is inserted at the target site; covering said population of hPSCs under an extracellular matrix comprising at least one neuronal differentiation driver to produce a sensory committed neural crest population; contacting said sensory committed neural crest population with at least one neuronal differentiation driver and at least one neurotrophic factor to produce a population of early sensory neurons (SNs); and contacting said population of early SNs with at least one neurotrophic factor to produce a population of mature hPSC-SNs expressing at least one of SN marker or one pan neuronal marker; isolating the cells expressing the at least one screenable, selectable marker, wherein said population of mature hPSC-SNs expressed said one target gene associated with nociceptive pain, chronic pain, pruriception a nociceptive- or pruriceptive- mediated condition, and wherein said population of mature hPSC-SNs responds to a nociceptive and pruriceptive stimulus.

Levels of expression of antibiotic resistance genes are increased up to six-fold by inserting a proteasome-targeting tag into transgenes expressed in eukaryotic cells. Various selectable marker proteins are combined with different destabilization domains, leading to up to 70% increase in transgene expression. The increase in expression varies highly depending on the engineered construct and the lines cells used. Increase in expression drives exosome loading of cargo proteins in some aspects. By increasing expression and by editing trafficking signals of cargo proteins, proteins that normally locate to the ER can be trafficked to exosomes. This disclosure discloses efficient exosome delivery of a wide variety of engineered proteins, including modified antigen proteins of SARS-CoV-2 and influenza, and other proteins such as a modified alpha galactosidase A, an extracellular domain of vascular endothelial growth factor fused to a constant region of a human immunoglobulin heavy chain, and modified trastuzumab heavy and light chains.

Dual vector systems and these comprehensive mammalian host cells are provided for the expression of multi-chain molecules, with each vector employing a different promoter controlling the expression of a selectable marker. The mammalian host cells can be used to generate a recombinant protein of interest in a mammalian cell culture.

Dual vector systems and mammalian host cells comprising these dual vector systems are provided, wherein each vector employs a different promoter that drives the expression of a selectable marker. A mammalian host cell can be used to produce a recombinant protein of interest in mammalian cell culture.

The invention provides a method for dynamically regulating and controlling the copy number of plasmids in saccharomyces cerevisiae based on RNA (Ribonucleic Acid) interference. According to the method, an orthogonal RNAi system is reconstructed, saccharomyces cerevisiae is used as a synthetic biological chassis with a dynamic programmable plasmid copy number, a heterologous RNAi mechanism from Ckstri saccharomyces cerevisiae is integrated, sequence-specific siRNA of a targeted plasmid coding selective marker is designed, a chemically induced gene dose control platform is established, and the gene dose control platform is used for controlling the gene dose. The dynamic regulation and control of the plasmid copy number are realized, so that the gene expression dosage is optimized, and the biosynthesis efficiency is improved. RNAi-mediated copy number regulation is applied to a carotenoid biosynthesis pathway, and compared with a static plasmid system, the lycopene titer is increased by 18.6 times. The method further enriches an anabolism regulation tool set, has huge potential in the aspect of improving the production performance of a microbial cell factory, and provides a new thought for metabolic engineering optimization in the field of synthetic biology.

Herein is reported a nucleic acid comprising in operably linked form a nucleic acid encoding a selection marker, a nucleic acid encoding a self-cleaving peptide sequence, and a nucleic acid encoding a proteinaceous protease inhibitor. Further reported are methods for the recombinant production of a heterologous polypeptide using said nucleic acid as well as a cell comprising said nucleic acid according to the invention. Likewise reported is the use of the nucleic acid according to the invention for increasing the amount of the recombinantly produced heterologous polypeptide by reducing protease cleavage.

Provided are methods for inserting a large gene / polynucleotide between about 4.5Kb and about 8Kb and lacking a selection marker into a target genomic position in a cell by delivering the gene / polynucleotide by two AAV vectors in the presence of a p53-binding protein 1 (53BP1) inhibitor and a DNA-dependent protein kinase catalytic subunit (DNA- PKcs) inhibitor. The disclosed methods improve large gene insertion by at least 10-fold and obviate the need for a selection marker in the vector design, thereby providing more room in the AAV vector for gene delivery.

The application discloses a dCAPS molecular marker for detecting a residual amount of a cucumber fruit myclobutanil and application thereof, and relates to the technical field of molecular markers.The dCAPS molecular marker is derived from a SNP at a position of 158 of a CsABCI19 gene promoter.The dCAPS molecular marker is applied to detection of a residual amount of a cucumber fruit myclobutanil.The method comprises the following steps: (1) extracting DNA; (2) performing a PCR reaction; and (3) performing Nco I enzyme cutting, so that two band types are obtained, one is 491 bp, corresponding to SNP A, and the other is 444 bp and 48 bp, corresponding to SNP G, and the two types are identified as a fruit low myclobutanil residual amount germplasm and a fruit high myclobutanil residual amount germplasm respectively.The verification result of the dCAPS molecular marker in the germplasm resource shows that the identification result of the molecular marker is consistent with the phenotype, and the dCAPS molecular marker can be used for rapidly identifying the cucumber germplasm resource with a low myclobutanil residual amount in a fruit, and is a good breeding auxiliary selection marker.

The present invention relates to a method for integrating one or more recombinant sites in a sole target gene of a cell genome comprising carrying out sequentially multiple site-specific recombination by using alternatively two different serine-integrases, wherein the selection marker is removed after each site-specific recombination by different transposases: An in vitro method for inserting at least one recombinant site in a target gene / locus from an isolated cell genome, said method comprising integrating a nucleotide sequence A into the target gene from an isolated cell genome by homologous recombination, wherein said nucleotide sequence A comprises a recombination cassette, flanked at 5′ and 3′ by target gene homology arms, and removing the selection marker by using the second transposase, thus obtaining a cell genome A comprising (i), (ii) and (iii) of nucleotide sequence A.

The invention discloses an SNP molecular marker related to cottonseed oil content and application of the SNP molecular marker, and belongs to the technical field of molecular marker development. The SNP molecular marker is located at the 43284845th site of an upland cotton genome D03 chromosome, the nucleotide is A or G, the upstream and downstream sequences are shown as SEQ ID NO.1, and the G allele is associated with the high oil content character. The SNP molecular marker can be directly applied to (1) identification and screening of high-oil cotton germplasm resources: high-oil materials are rapidly screened from existing varieties or strains; (2) cotton molecular breeding practice: as an effective selection marker, the cotton molecular marker is used for guiding the breeding of filial generations, and by selecting individuals carrying G alleles (especially GG type), the directionality and success rate of breeding new varieties / lines of high-oil cotton are remarkably improved. The method has very important practical application value.

The present invention relates to a method for editing the genome of a cell, such as a yeast cell. The method of the present invention requires the cell to be contacted with at least one ribonucleoprotein, at least one donor-DNA construct and a selectable marker such that they are introduced into the cell. The present invention is especially suitable for multiplex genome editing of cells such as yeast cells. The current invention further relates to a composition, a cell obtainable by the method of the invention and a method for the production of a compound of interest.

The disclosure provides, for example, a method of making a composition comprising purified DNA molecules. In some embodiments, the method comprises contacting a solution comprising a plurality of circular, double-stranded DNA molecules to a hydrophobic interaction chromatography matrix. In some embodiments, the circular, double-stranded DNA molecules lack one or both of a bacterial origin of replication and a selectable marker.

The present invention provides for the production of improved viral and non-viral nanoplasmid vectors. Specifically, the invention provides an antibiotic-label-free covalent closed circular recombinant DNA molecule comprising: a. An antibiotic-label-free insert comprising a structured DNA sequence selected from the group consisting of a reverse repeat, a forward repeat, a homopolymerized repeat and an eukaryotic origin of replication; b. A Pol III-dependent origin of replication comprising an R6K [gamma] origin of replication having at least 95% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 18; and c. An RNA-OUT RNA selection marker, which comprises an RNA-IN modulating RNA-OUT RNA having at least 95% sequence identity with SEQ ID NO: 6, and a method for detecting the RNA-OUT RNA.

The invention provides a DNA marker plasmid as well as a combination system and application thereof, and relates to the field of molecular biology. The DNA marker plasmid is formed by connecting a linearized skeleton sequence and an insertion fragment sequence; the linearized skeleton sequence is derived from a reference plasmid from which a lactose operon sequence is deleted, and a selectable marker gene of the linearized skeleton sequence has a preset recognition site of restriction enzyme; the insertion fragment sequence is formed by connecting a plurality of DNA fragments in series, and recognition sites of restriction endonuclease are arranged between the adjacent DNA fragments. According to the plasmid, a restriction enzyme cutting site is introduced into a selective marker, so that a skeleton is directly used as a characteristic band, and the complete utilization of a sequence is realized. Waste and purification are avoided after enzyme digestion, the cost is remarkably reduced, and the preparation efficiency is improved.

The present invention provides an isolated nucleic acid encoding a reporter fusion protein comprising a selection marker, a protease recognition sequence and a degron.

The present invention provides host cells and vector systems that allow precise tuning of expression of selectable marker expression and increased integrated copy number of product genes so that host cell growth and expression of desired proteins or nucleic acid products from the host cell can be optimized.

The present disclosure provides a host cell for optogenetic protein production, comprising: a first sequence comprising a first promoter operably linked to a gene of interest; a second sequence comprising a second promoter operably linked to a gene encoding a light-sensitive transcription factor, wherein the light-sensitive transcription factor is configured to induce expression of the gene of interest upon activation by light; and a first selection marker. The gene of interest, first promoter, second gene, second promoter, and first selection marker are either integrated at a first locus, or the gene of interest, first promoter, and first selection marker are integrated at the first locus while the second gene, second promoter, and a second selection marker are integrated at a second locus. The disclosure also provides systems and methods for optogenetic protein production using the host cell.

This invention relates to a cell line containing a partially or completely inactivated endogenous dihydroorotate dehydrogenase (DHODH) gene and its use in producing recombinant proteins.

The invention relates to a modified mammalian glutamine synthetase comprising a mutation at amino acid position 10 and / or 298 in a mammalian glutamine synthetase, wherein the mutation is selected from the group consisting of R298K, N10S, N10T and N10Q and to an expression vector, a nucleic acid and a eukaryotic host cell encoding said modified mammalian glutamine synthetase and to its use as a selection marker. The invention further relates to methods for preparing stable cell lines, or for producing a protein of interest using said modified mammalian glutamine synthetase for selection.

The present invention provides a novel marker for genetic recombination in Starmerella bombicola and a method for using the same. The present invention also provides a Starmerella bombicola mutant strain in which the expression of an orotate phosphoribosyltransferase-like protein or a protein equivalent thereto is suppressed or inactivated, a uracil-requiring selection marker consisting of the gene, and a method for producing a Starmerella bombicola transformant using the selection marker.

A marker for gene recombination of Starmerella bombicola and a method for using the same. The present invention provides an adenine auxotrophic selection marker consisting of a gene encoding a phospho

The invention provides bidirectional expression vectors comprising Chinese hamster ovary elongation factor 1-α (CHEF1) transcriptional regulatory DNA elements, a gene of interest (GOI), a minimal cytomegalovirus (minCMV) and a selectable marker (SM) and / or a human adenovirus tripartite leader (AdTPL) sequence. The invention also provides method for increasing heterologous protein expression in a host cell comprising culturing the host cell the bidirectional expression vector(s).

A lentivlral vector for real-time monitoring of cell state which includes a regulatory element that regulates expression of a selection marker, wherein the regulatory element comprises cell-state-specific histone H3K27-acetylation marks and / or chromatin accessibility. Methods of identifying a cell- state-specific regulatory element in a cell; optically screening a compound for the ability to modify cell identity; in vivo labeling normal and / or diseased cells at a specific developmental stage or a specific stage of disease; and real- time monitoring of transcription of a cell exposed to a stress using the lentiviral vector.

A method of differentiating nonhuman primate induced pluripotent stem cells (iPSCs) to oligodendrocytes is provided. The method may include providing a modified vector comprising a piggyBac vector backbone that expresses one or more differentiation factor genes, including SOX10, OLIG2, and NKX6-2. The piggyBac vector backbone may comprise one or more terminal inverted repeats and one or more transposase recognition sites configured to interact with a piggyBac transposase to mediate genomic integration at TTAA or noncanonical target sequences. The vector may further include a selectable marker and a promoter for regulated expression. The modified vector may be introduced into nonhuman primate iPSCs, for example from Macaca fascicularis, by electroporation and used in a feeder-free differentiation workflow. The resulting differentiated cells may be identified by expression of oligodendrocyte-associated markers including O4, O1, PDGFRA, MOG, GALC, CNPase, and myelin basic protein (MBP).

Provided are a novel genetic recombination marker of Starmerella bombicola, and a method for using the same. An adenine-requirement selectable marker comprising a gene encoding a phosphoribosylglycinamide formyltransferase-like protein or an equivalent gene. A mutant strain of Starmerella bombicola in wich the expression of a phosphoribosylglycinamide formyltransferase-like protein or an equivalent protein is suppressed or inactivated.

Disclosed herein are novel selectable markers and uses thereof. Specifically, provided herein is a nucleotide sequence encoding glutamine synthetase (GS) derived from Taokuo bear, ostrich, snake, pigeon, chicken, gerbil, or bat as a selectable marker for identifying genomic loci with high transcriptional activity or host cells with high productivity of a protein of interest, and / or a use for accelerating the identification process. Related screening methods, production methods and expression systems are also included.

To provide a method for producing a human embryonic kidney (HEK) cell line that does not express functional endogenous dihydrofolate reductase (DHFR) and does not express functional endogenous glutamine synthetase (GS).SOLUTION: The present invention provides a method comprising the steps of: (a) mutating or knocking out the endogenous DHFR gene and the GS gene in a HEK cell line; (b) stably or transiently transfecting the HEK cell line with an AAV vector genome comprising a first heterologous nucleic acid sequence encoding a therapeutic protein or an inhibitory nucleic acid sequence and a first selectable marker; and (c) stably or transiently transfecting the HEK cell line with a second heterologous nucleic acid sequence and a second selectable marker.SELECTED DRAWING: Figure 2